Human Colorectal Carcinoma Tissues Research Articles

Relationship between expression of laminin and pathological features in human colorectal carcinoma.

AIM:To study the expression and significance of laminin in human colorectal carcinoma.METHODS:Using the monoclonal antibody to laminin and streptavidin-peroxidase immunohistochemical method, the expression of laminin in 63 cases of human colorectal carcinoma tissues was determined.RESULTS:In normal marge intestinal mucosa adjacent to carcinoma, laminin was largely restricted to basement membrane in continuous linear pattern. In contrast, human colorectal carcinomas exhibited a progressive loss of an intact basement membrane that was correlated with decreasing differentiation degree.Well and moderately differentiated tumors exhibited a thin basement membrane with intermittent disruptions, and poorly differentiated tumors exhibited no areas of intact basement membrane. An association was found between lack of basement membrane laminin immunohistochemical staining in colorectal carcinoma and poorly differentiated tumor (P < 0.01).CONCLUSION:Immunohistochemical staining for laminin could provide a very useful indexfor the determination of the differentiation degree of colorectal carcinoma.

Reduced messenger RNA expression level of p21CIP1 in human colorectal carcinoma tissues and its association with p53 gene mutation

Reduced messenger RNA expression level of p21CIP1 in human colorectal carcinoma tissues and its association with p53 gene mutation

Reduced messenger RNA expression level of p21 CIP1 in human colorectal carcinoma tissues and its association with p53 gene mutation.

p21 Cip1 was first isolated as one of the cyclin-dependent kinase (Cdk) interacting proteins induced by wild-type p53 gene product, and it appears to play an essential regulatory role in the control of cell proliferation as a potent, tight-binding inhibitor of cyclin-Cdk complex that blocks the G1/S transition of the cell cycle. We have now examined the p21 Cip1 mRNA expression levels in 16 surgically excised human colorectal tumor and non-tumor tissues by Northern-blot analysis with reference to the identification of p53 gene mutations. p53 gene mutations were detected in 6 tumor tissues but not in the other 10 tissues by the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method and following direct sequencing. The mean p21 Cip1 mRNA expression level in tumor tissues was significantly suppressed compared to that of non-tumor tissues, irrespective of p53 gene mutations. In p53 gene mutation-detected cases, the mean expression level of p21 Cip1 mRNAs of tumor tissues was about 60% of that of cases without p53 gene mutation. Moreover, the relative mRNA expression levels of p21 Cip1 significantly decreased as the pathohistological stages progressed by Dukes' staging system, while in patients with liver metastasis these levels were significantly suppressed compared to those of patients without organ metastasis. These results indicate that reduced expression of p21 Cip1 mRNA is critical for growth activity and malignant potential of human colorectal carcinoma, and that the decrease in p21 Cip1 mRNA level is due to p53 gene mutation as well as other mechanisms during human colorectal carcinogenesis.

Different sialyltransferase activities in human colorectal carcinoma cells from surgical specimens detected by specific glycoprotein and glycolipid acceptors.

The amount and type of sialylation of tumor cell membranes depends on the activity of a number of different sialyltransferase enzymes. For the detection of specific activities in human colorectal carcinoma tissue several glycoprotein and glycolipid acceptors were used: desialylated fetuin, alpha 1-acid glycoprotein, beta 2-glycoprotein I, ovine submaxillaris mucin, and the gangliosides GM1, GM2, GM3 and GD1a. Because of their possible relevance for metastasis, precursors of Le(a) and Le(x) antigens, too, were employed, namely neoglycolipids produced by coupling LcOse4 or NeoLcOse4 oligosaccharides to L-alpha-phosphatidyl-ethanol-amine-dipalmitoyl. Our data indicate that human colorectal tumor tissue contains two highly active sialyltransferase enzymes, which are only weakly expressed in normal mucosa. These are a N-glycan-specific alpha 2,6-sialyltransferase, which was significantly increased in metastasizing tumors, and a Gal beta 1,3Gal-NAc-specific sialyltransferase, which was increased in tumors of early stages. A shift to enhanced alpha 2,6-sialylation of membrane glycoproteins during carcinogenesis was demonstrated by lectin ELISA analysis of magneto-bead separated tumor cells. Quantitative determination of specific sialyltransferase activities may be a sensitive tool for detection and monitoring of colon carcinoma.

Prevalence of G-to-T transversions among K-ras oncogene mutations in human colorectal tumors in Yugoslavia.

Human colorectal carcinoma tissue sampled from 37 patients, routinely graded into Dukes' stages A, B and C and histologically examined for the level of differentiation, were analyzed for the presence of point mutations in the K-ras oncogene. Seventeen cases out of the 37 analyzed were found to have a mutation in either the 12th or the 13th codon of the K-ras gene, giving an overall frequency of mutation of 46%. The incidence of mutations in Dukes' stages A, B and C was 33, 46 and 58% respectively. Although the frequency of mutation appears to be similar to that reported for the USA population, the spectrum of point mutations in codons 12 and 13 of the K-ras gene in the Yugoslav population appears to differ significantly. G-to-T transversions make up 77% of all mutations present, with the distribution as follows: 18% at the first base and 59% at the second base of codons 12 and 13. G-to-A transitions at the second base is the only other mutation identified, occurring mainly in codon 13 in colorectal tumors of all 3 stages.

Plasminogen activators and plasminogen activator inhibitors in human colorectal carcinoma tissues are not expressed by the tumour cells

Plasminogen activators (PA) have been implicated with the degradation of extracellular matrix during the invasive growth of metastasising tumour cells. The significance of PA expression in tumour cells for the in vivo growth of malignant tumours is still a matter of debate. We, therefore, performed immunohistological studies on human colon tumours using monoclonal antibodies against urokinase- (u-PA) and tissue-type plasminogen activator (t-PA) as well as against plasminogen activator inhibitors 1 and 2 (PAI-1, PAI-2). Normal colorectal mucosa of seven samples was negative for all four constituents of the PA system. Tumour epithelium of 64 colorectal carcinomas and 10 liver metastases was consistently negative for both, PA and their inhibitors. However, two of four human colon carcinoma cell lines weakly expressed u-PA, PAI-1 and PAI-2. Intestinal dendritic or fibroblast-like cells within the tumour tissue strongly expressed u-PA and, at a lower level, also t-PA, PAI-1 and PAI-2. Vascular endothelial cells were weakly positive for all components of the PA system in colon carcinoma. Our findings indicate that colon carcinoma cells in their natural environment do not express constituents of the PA system. PA activity, previously found in colon carcinoma tissue, is most likely derived from interstitial cells.

Separation of tumor cells from a suspension of dissociated human colorectal carcinoma tissue by means of monoclonal antibody-coated magnetic beads

Dissociation of colorectal tumor tissue directly obtained at surgery yields a cell suspension which contains tumor cells, leukocytes and erythrocytes. Coupling of a murine monoclonal antibody, HEA125, specific for human epithelial cells, to magnetic beads permits positive selection of a population containing essentially only tumor cells. Removal of cells from the beads is possible by papain treatment. Negative selection of tumor cells by means of beads coated with a leukocyte binding antibody offers no advantage since dead cells and erythrocytes remain.

Direct isolation of inoue‐melnick virus from human colorectal carcinoma tissues but not from stomach carcinoma tissues

Inoue-Melnick virus (IMV) was isolated directly from all of the seven human colorectal adenocarcinoma tissues studied, whereas IMV was not isolated from five human stomach carcinoma tissues. Seven isolates from the Japanese colorectal carcinoma tissues were identified as type 1 IMV by the neutralization test and were sensitive to phosphonoacetic acid, a growth inhibitor of IMV. Furthermore, type 1 IMV-antibody was detected in six of seven sera of the patients whose colorectal carcinoma tissues were positive for virus isolation. In contrast, IMV-antibody was not detected in the five sera of the patients whose stomach carcinoma tissues were tested and found negative for IMV.

Occurrence of an active form of gelatinase in human gastric and colorectal carcinoma tissues

Profiles of gelatinases released during 4 h incubation of explants from human carcinoma tissues (15 gastric and 15 colorectal carcinomas) in serum-free RPMI 1640 medium were compared with those of corresponding normal tissue specimens obtained 2 and 10 cm apart from tumor periphery. All the culture supernatants of both tumor and normal tissues contained gelatinase species of 200, 130, 92 and 72 kDa as detected by zymography with gelatin as a substrate. Besides these gelatinase species, all carcinoma tissue culture supernatants had an additional 66-kDa gelatinase, an active from of the 72-kDa gelatinase. This gelatinase was not detectable in the normal tissues 10 cm apart from the tumor mass, with an exceptional case of gastric carcinoma of Borrmann type 4. These results suggest that the presence of active form of gelatinase might be one of the characteristic properties of these malignant human tumors.

Verapamil increases the sensitivity of primary human colorectal carcinoma tissue to vincristine

Verapamil increases the sensitivity of primary human colorectal carcinoma tissue to vincristine